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Win1 phosphorylates and activates Wis1 in vitro. (A) Win1 phosphorylates Wis1. In vitro kinase activity of the catalytic domain of Win1 purified from S. pombe was tested with recombinant GST-Wis1(K349R) as a substrate. (B) Win1 activates Wis1. Wis1 proteins were preincubated with Win1 proteins in the presence of unlabeled ATP. Then, Wis1 kinase activity was tested by assaying incorporation of radioactive phosphate into GST-Spc1 after adding γ-labeled ATP.
